Aguayo Vasti, Valdes Bianca, Espino Ana M
Laboratory of Immunology and Molecular Parasitology, Department of Microbiology, University of Puerto Rico, School of Medicine, San Juan, PR 00936-5067, United States.
Laboratory of Immunology and Molecular Parasitology, Department of Microbiology, University of Puerto Rico, School of Medicine, San Juan, PR 00936-5067, United States.
Acta Trop. 2018 Oct;186:41-49. doi: 10.1016/j.actatropica.2018.07.002. Epub 2018 Jul 7.
Due to the unsatisfactory performance of parasitological diagnosis of human fascioliasis; the use of immunodiagnosis based on the detection of anti-Fasciola antibodies is traditionally used as a diagnostic alternative using total or purified parasite excretory-secretory products (ESPs). Glutathione S-transferase (GST) protein, one of the F. hepatica ESP components, possesses well-known roles in the detoxification of xenobiotic and endogenously derived toxins within the host bile environment. GST has shown to be a good target for vaccine or drug development against fascioliasis. The current study aimed to evaluate the potential of GST protein purified from a soluble crude extract of adult flukes as an antigen for serodiagnosis of chronic human fascioliasis by indirect ELISA. The study included a panel of 116 serum samples collected from individuals with confirmed fascioliasis, individuals carrying heterologous parasitic infections and healthy subjects. The parasitological examination was used as gold standard and a previously optimized ESP-ELISA was used to compare the performance of the GST-ELISA method. Results demonstrated that GST-ELISA is 94.3% sensitive, 80.2% specific and exhibits a moderate positive correlation (r = 0.555) and substantial agreement (k = 0.786) with the results obtained with the ESP-ELISA method. Moreover, because no sera from patients with early F. hepatica infection were available, GST-ELISA was then tested with sera from rabbits experimentally infected with F. hepatica metacercariae. The assay was able to detect anti-Fasciola antibodies as early as the 3 week of infection (p < 0.0001) with peaks at 4 and 10 week post-infection.
由于人体肝片吸虫病的寄生虫学诊断效果不尽人意;传统上,基于检测抗肝片吸虫抗体的免疫诊断被用作一种诊断替代方法,使用的是寄生虫全虫或纯化的排泄分泌产物(ESPs)。谷胱甘肽S-转移酶(GST)蛋白是肝片吸虫ESPs的组成成分之一,在宿主胆汁环境中对外源和内源性毒素的解毒过程中发挥着众所周知的作用。GST已被证明是抗肝片吸虫病疫苗或药物开发的一个良好靶点。本研究旨在评估从成年吸虫可溶性粗提物中纯化的GST蛋白作为慢性人体肝片吸虫病血清学诊断抗原的潜力。该研究纳入了一组116份血清样本,这些样本来自确诊为肝片吸虫病的个体、携带异源寄生虫感染的个体以及健康受试者。寄生虫学检查被用作金标准,并且使用先前优化的ESPs-ELISA来比较GST-ELISA方法的性能。结果表明,GST-ELISA的敏感性为94.3%,特异性为80.2%,与ESPs-ELISA方法获得的结果呈现中度正相关(r = 0.555)和高度一致性(k = 0.786)。此外,由于没有早期肝片吸虫感染患者的血清,随后用实验感染肝片吸虫囊蚴的家兔血清对GST-ELISA进行了检测。该检测方法能够在感染后第3周就检测到抗肝片吸虫抗体(p < 0.0001),在感染后第4周和第10周出现峰值。